目的 探讨大鼠心肌缺血再灌注损伤(MIRI)前、后使用脂氧素A4(Lxa4a)对心肌组织Na+-K+-ATP酶的影响及意义.方法 72只SD大鼠随机分成假手术一组(C1组)、假手术二组(C2组)、MIRI一组(I/R1组)、MIRI二组(I/R2组)、MIRI前用药组(LX1组)、MIRI后用药组(Lx2组),每组12只.建立大鼠MIRI模型,各组于开胸前取血(T1)、实验结束后取血(T2)测血清cTnI浓度.检测Na+-K+-ATP酶蛋白、mRNA的表达及活性改变;同时检测心肌细胞凋亡率,测定SOD活性、MDA含量,电镜下观察各组超微结构的改变.结果 I/R1、LX1与C1相比,I/R2、LX2与C2相比,Na+-K+-ATP酶蛋白(α1:0.19±0.03,0.29±0.05比0.46±0.06,0.21±0.03,0.36±0.05比0.48±0.07,α2:0.20±0.02,0.32±0.04比0.54±0.05,0.23±0.03,0.38±0.04比0.56±0.06,α3:0.24±0.03,0.35±0.04比0.61±0.04,0.26±0.03,0.41±0.04比0.59±0.04,β1:0.22±0.03,0.33±0.04比0.56±0.04,0.24±0.03,0.39±0.04比0.54±0.04),mRNA的表达下降[(0.21±0.07)×106,(0.55 ±0.28)×106比(5.73±1.97)×106,(0.20±0.09)×106,(1.78±0.86)×106比(6.21±2.22)×106],活性(mmol/gprot)降低(0.73±0.04,0.95±0.11比1.38±0.15,0.75±0.04,1.19±0.12比1.37±0.13);血清cTnI浓度(ng/L) (T2)(580.93±43.92,292.95±21.99比151.53±14.00,592.80±60.83,207.13±28.54比155.18±14.92),凋亡指数(52.52±6.36,34.55±5.65比7.48±2.09,54.75±6.29,26.53±4.60比8.54±2.76)以及SOD活性(U/mgprot) (196.84±31.82,293.59±43.10比102.55±24.77,202.48±35.40,411.71±60.44比113.36±26.95) 、MDA含量(nmol/gprot)(2402.94± 438.56,1548.57±238.08比472.48±190.77,2422.28±517.74,1055.48±237.44比483.26±172.05)显著增高(P＜0.05). LX1与I/R1,LX2与I/R2相比,Na+-K+-ATP酶蛋白(α1:0.29±0.05比0.19±0.0 3,0.36±0.05比0.21±0.03,α2:0.32±0.04比0.20±0.02,0.38±0.04比0.23±0.03,α3:0.35±0.04比0.24±0.03,0.41±0.04比0.26±0.03,β1:0.33±0.04比0.22±0.03,0.39±0.04比0.24±0.03) 、mRNA的表达[(0.55±0.28)×106比(0.21±0.07)×106,(1.78±0.86)×106比0.20±0.09)×106]明显增加,Na+-K+-ATP酶活性(mmol/gprot)(0.95±0.11比0.73±0.04,1.19±0.12比0.75±0.04)与SOD的活性(U/mgprot) (293.59±43.10比196.84±31.82,411.71±60.44比202.48±35.40)明显增强;同时凋亡指数(34.55±5.65比52.52±6.36,26.53±4.60比54.75±6.29)、血清cTnI浓度(ng/L)(T2)(292.95±21.99比580.93±43.92,207.13±28.54比592.80±60.83) 、MDA含量(nmol/gprot)(1548.57±238.08比2402.94±438.56,1055.48±237.44比2422.28±517.74)也明显降低(P＜0.05).LXA4对缺血再灌注后心肌超微结构有明显的保护作用.结论 在MIRI前、后应用LXA4能明显减轻心肌氧化损伤,上调Na+-K+-ATP酶蛋白、mRNA的表达,增强Na+-K+-ATP酶的活性,保护心肌超微结构,减少细胞凋亡,发挥其心肌保护作用.
目的 探討大鼠心肌缺血再灌註損傷(MIRI)前、後使用脂氧素A4(Lxa4a)對心肌組織Na+-K+-ATP酶的影響及意義.方法 72隻SD大鼠隨機分成假手術一組(C1組)、假手術二組(C2組)、MIRI一組(I/R1組)、MIRI二組(I/R2組)、MIRI前用藥組(LX1組)、MIRI後用藥組(Lx2組),每組12隻.建立大鼠MIRI模型,各組于開胸前取血(T1)、實驗結束後取血(T2)測血清cTnI濃度.檢測Na+-K+-ATP酶蛋白、mRNA的錶達及活性改變;同時檢測心肌細胞凋亡率,測定SOD活性、MDA含量,電鏡下觀察各組超微結構的改變.結果 I/R1、LX1與C1相比,I/R2、LX2與C2相比,Na+-K+-ATP酶蛋白(α1:0.19±0.03,0.29±0.05比0.46±0.06,0.21±0.03,0.36±0.05比0.48±0.07,α2:0.20±0.02,0.32±0.04比0.54±0.05,0.23±0.03,0.38±0.04比0.56±0.06,α3:0.24±0.03,0.35±0.04比0.61±0.04,0.26±0.03,0.41±0.04比0.59±0.04,β1:0.22±0.03,0.33±0.04比0.56±0.04,0.24±0.03,0.39±0.04比0.54±0.04),mRNA的錶達下降[(0.21±0.07)×106,(0.55 ±0.28)×106比(5.73±1.97)×106,(0.20±0.09)×106,(1.78±0.86)×106比(6.21±2.22)×106],活性(mmol/gprot)降低(0.73±0.04,0.95±0.11比1.38±0.15,0.75±0.04,1.19±0.12比1.37±0.13);血清cTnI濃度(ng/L) (T2)(580.93±43.92,292.95±21.99比151.53±14.00,592.80±60.83,207.13±28.54比155.18±14.92),凋亡指數(52.52±6.36,34.55±5.65比7.48±2.09,54.75±6.29,26.53±4.60比8.54±2.76)以及SOD活性(U/mgprot) (196.84±31.82,293.59±43.10比102.55±24.77,202.48±35.40,411.71±60.44比113.36±26.95) 、MDA含量(nmol/gprot)(2402.94± 438.56,1548.57±238.08比472.48±190.77,2422.28±517.74,1055.48±237.44比483.26±172.05)顯著增高(P＜0.05). LX1與I/R1,LX2與I/R2相比,Na+-K+-ATP酶蛋白(α1:0.29±0.05比0.19±0.0 3,0.36±0.05比0.21±0.03,α2:0.32±0.04比0.20±0.02,0.38±0.04比0.23±0.03,α3:0.35±0.04比0.24±0.03,0.41±0.04比0.26±0.03,β1:0.33±0.04比0.22±0.03,0.39±0.04比0.24±0.03) 、mRNA的錶達[(0.55±0.28)×106比(0.21±0.07)×106,(1.78±0.86)×106比0.20±0.09)×106]明顯增加,Na+-K+-ATP酶活性(mmol/gprot)(0.95±0.11比0.73±0.04,1.19±0.12比0.75±0.04)與SOD的活性(U/mgprot) (293.59±43.10比196.84±31.82,411.71±60.44比202.48±35.40)明顯增彊;同時凋亡指數(34.55±5.65比52.52±6.36,26.53±4.60比54.75±6.29)、血清cTnI濃度(ng/L)(T2)(292.95±21.99比580.93±43.92,207.13±28.54比592.80±60.83) 、MDA含量(nmol/gprot)(1548.57±238.08比2402.94±438.56,1055.48±237.44比2422.28±517.74)也明顯降低(P＜0.05).LXA4對缺血再灌註後心肌超微結構有明顯的保護作用.結論 在MIRI前、後應用LXA4能明顯減輕心肌氧化損傷,上調Na+-K+-ATP酶蛋白、mRNA的錶達,增彊Na+-K+-ATP酶的活性,保護心肌超微結構,減少細胞凋亡,髮揮其心肌保護作用.
목적 탐토대서심기결혈재관주손상(MIRI)전、후사용지양소A4(Lxa4a)대심기조직Na+-K+-ATP매적영향급의의.방법 72지SD대서수궤분성가수술일조(C1조)、가수술이조(C2조)、MIRI일조(I/R1조)、MIRI이조(I/R2조)、MIRI전용약조(LX1조)、MIRI후용약조(Lx2조),매조12지.건립대서MIRI모형,각조우개흉전취혈(T1)、실험결속후취혈(T2)측혈청cTnI농도.검측Na+-K+-ATP매단백、mRNA적표체급활성개변;동시검측심기세포조망솔,측정SOD활성、MDA함량,전경하관찰각조초미결구적개변.결과 I/R1、LX1여C1상비,I/R2、LX2여C2상비,Na+-K+-ATP매단백(α1:0.19±0.03,0.29±0.05비0.46±0.06,0.21±0.03,0.36±0.05비0.48±0.07,α2:0.20±0.02,0.32±0.04비0.54±0.05,0.23±0.03,0.38±0.04비0.56±0.06,α3:0.24±0.03,0.35±0.04비0.61±0.04,0.26±0.03,0.41±0.04비0.59±0.04,β1:0.22±0.03,0.33±0.04비0.56±0.04,0.24±0.03,0.39±0.04비0.54±0.04),mRNA적표체하강[(0.21±0.07)×106,(0.55 ±0.28)×106비(5.73±1.97)×106,(0.20±0.09)×106,(1.78±0.86)×106비(6.21±2.22)×106],활성(mmol/gprot)강저(0.73±0.04,0.95±0.11비1.38±0.15,0.75±0.04,1.19±0.12비1.37±0.13);혈청cTnI농도(ng/L) (T2)(580.93±43.92,292.95±21.99비151.53±14.00,592.80±60.83,207.13±28.54비155.18±14.92),조망지수(52.52±6.36,34.55±5.65비7.48±2.09,54.75±6.29,26.53±4.60비8.54±2.76)이급SOD활성(U/mgprot) (196.84±31.82,293.59±43.10비102.55±24.77,202.48±35.40,411.71±60.44비113.36±26.95) 、MDA함량(nmol/gprot)(2402.94± 438.56,1548.57±238.08비472.48±190.77,2422.28±517.74,1055.48±237.44비483.26±172.05)현저증고(P＜0.05). LX1여I/R1,LX2여I/R2상비,Na+-K+-ATP매단백(α1:0.29±0.05비0.19±0.0 3,0.36±0.05비0.21±0.03,α2:0.32±0.04비0.20±0.02,0.38±0.04비0.23±0.03,α3:0.35±0.04비0.24±0.03,0.41±0.04비0.26±0.03,β1:0.33±0.04비0.22±0.03,0.39±0.04비0.24±0.03) 、mRNA적표체[(0.55±0.28)×106비(0.21±0.07)×106,(1.78±0.86)×106비0.20±0.09)×106]명현증가,Na+-K+-ATP매활성(mmol/gprot)(0.95±0.11비0.73±0.04,1.19±0.12비0.75±0.04)여SOD적활성(U/mgprot) (293.59±43.10비196.84±31.82,411.71±60.44비202.48±35.40)명현증강;동시조망지수(34.55±5.65비52.52±6.36,26.53±4.60비54.75±6.29)、혈청cTnI농도(ng/L)(T2)(292.95±21.99비580.93±43.92,207.13±28.54비592.80±60.83) 、MDA함량(nmol/gprot)(1548.57±238.08비2402.94±438.56,1055.48±237.44비2422.28±517.74)야명현강저(P＜0.05).LXA4대결혈재관주후심기초미결구유명현적보호작용.결론 재MIRI전、후응용LXA4능명현감경심기양화손상,상조Na+-K+-ATP매단백、mRNA적표체,증강Na+-K+-ATP매적활성,보호심기초미결구,감소세포조망,발휘기심기보호작용.
Objective To investigate the influence and Significance of LipoxinA4 (LXA4) on myocardial Na+-K+-ATP enzyme in rats undergoing ischemia reperfusion injury.Methods Seventytwo SD rats were randomly divided into six groups (n =12):sham operation group one (group C1) ;sham operation group two (group C2) ; MIRI group one (group I/R1) ; MIRI group two (group I/R2) ; pre-MIRI treatment group one (group LX1) ; post-MIRI treatment group two (group LX2).The rat model of MIRI was established,the serum cTnI concentrations were measured in each group before chest opening (T1) and at the end of the experiment (T2).The expression and activity change of Na+-K+-ATP enzyme protein,mRNA were detected.At the same time,myocardial cell apoptosis was detected with TUNEL method,the SOD activation and MDA content were checked,while the ultrastructural changes of cardiac muscle were observed under electron microscope.Results In group I/R1,LX1 (compared with group C1) and group I/R2,LX2 (compared with group C2),the expression levels of Na+-K+-ATP protein (α1:0.19 ± 0.03,0.29 ± 0.05 to 0.46 ± 0.06,0.21 ± 0.03,0.36 ±0.05 to 0.48 ± 0.07,α2:0.20 ± 0.02,0.32 ± 0.04 to 0.54 ± 0.05,0.23 ± 0.03,0.38 ± 0.04 to 0.56 ±0.06,α3:0.24 ± 0.03,0.35 ± 0.04 to 0.61 ± 0.04,0.26 ± 0.03,0.41 ± 0.04 to 0.59 ± 0.04,β1:0.22 ±0.03,0.33 ± 0.04 to 0.56 ± 0.04,0.24 ± 0.03,0.39 ± 0.04 to 0.54 ± 0.04) and mRNA((0.21 ± 0.07) × 106,(0.55 ± 0.28) × 10to (5.73 ± 1.97) × 106,(0.20 ± 0.09) × 106,(1.78 ± 0.86) × 106 to (6.21 ±2.22) × 106).The activities of Na+-K+-ATP (mmol/gprot) were decreased respectively.Serum cTnI concentrations (T2),the apoptotic index and SOD activation (U/mgprot),MDA content (nmol/gprot) all increased significantly (P＜0.05).In group LX1 (comPared with group I/R1) and group LX2 (compared with group I/R2),the expression of Na+-K+-ATP protein,mRNA and activity of Na+-K+-ATP (mmol/gprot),SOD (U/mgprot) were raised significantly,while the serum cTnI concentrations (T2) (ng/L),MDA content (nmol/gprot) and apoptotic index were decreased significantly (P ＜ 0.05).LXA4 had obvious protective effect on myocardial ultrastructure after ischemia reperfusion.Conclusions Before and after MIRI,application of LXA4 could significantly attenuate myocardial oxidative injury,upregulate the expression of Na+-K+-ATP protein and mRNA,enhance the activity of Na+-K+-ATP enzyme.LXA4 plays its role in myocardial protection via the protection of myocardial ultrastructure and reduced apoptosis.
